A novel technique for quantifying changes in vascular density, endothelial cell proliferation and protein expression in response to modulators of angiogenesis using the chick chorioallantoic membrane (CAM) assay.

Bottom Line:
Reliable quantitative evaluation of molecular pathways is critical for both drug discovery and treatment monitoring.This improved CAM assay can correlate changes in vascular density with changes seen on a molecular level.We expect that these described modifications will result in a single in vivo assay system, which will improve the ability to investigate molecular mechanisms underlying the angiogenic response.

ABSTRACTReliable quantitative evaluation of molecular pathways is critical for both drug discovery and treatment monitoring. We have modified the CAM assay to quantitatively measure vascular density, endothelial proliferation, and changes in protein expression in response to anti-angiogenic and pro-angiogenic agents. This improved CAM assay can correlate changes in vascular density with changes seen on a molecular level. We expect that these described modifications will result in a single in vivo assay system, which will improve the ability to investigate molecular mechanisms underlying the angiogenic response.

A novel technique for quantifying changes in vascular density, endothelial cell proliferation and protein expression in response to modulators of angiogenesis using the chick chorioallantoic membrane (CAM) assay.

Mentions:
The amount of alpha v beta3 staining was greatest in stimulated CAM disks treated with bFGF. There was a significant decrease in the amount of alpha v beta3 in CAM disks after systemic treatment with fumagillin (P = 0.01, Student's T test). Treatment with LM609 did not significantly reduce the expression of alpha v beta 3 (P = 0.1376, Alternate Welch t test) although the values trended lower than control. Finally, there was a significant decrease in the amount of cleaved caspase 3 in CAM disks after systemic treatment with fumagillin (P = 0.007, Student's t test) and LM609 (P = 0.014, Student's t test, Figure 5B). This finding is consistent with previous data demonstrating a relationship between fumagillin treatment and inhibition of cleaved caspase-3 [14].

Bottom Line:
Reliable quantitative evaluation of molecular pathways is critical for both drug discovery and treatment monitoring.This improved CAM assay can correlate changes in vascular density with changes seen on a molecular level.We expect that these described modifications will result in a single in vivo assay system, which will improve the ability to investigate molecular mechanisms underlying the angiogenic response.

ABSTRACTReliable quantitative evaluation of molecular pathways is critical for both drug discovery and treatment monitoring. We have modified the CAM assay to quantitatively measure vascular density, endothelial proliferation, and changes in protein expression in response to anti-angiogenic and pro-angiogenic agents. This improved CAM assay can correlate changes in vascular density with changes seen on a molecular level. We expect that these described modifications will result in a single in vivo assay system, which will improve the ability to investigate molecular mechanisms underlying the angiogenic response.